MicroRNA‑224 is downregulated in mucinous cystic neoplasms of the pancreas and may regulate tumorigenesis by targeting Jagged1

Zhang,Beibei; Guo,Xiaorong; Zhang,Jingxi; Liu,Xiao; Zhan,Xianbao; Li,Zhaoshen

doi:10.3892/mmr.2014.2658

MicroRNA‑224 is downregulated in mucinous cystic neoplasms of the pancreas and may regulate tumorigenesis by targeting Jagged1

Authors:
- Beibei Zhang
- Xiaorong Guo
- Jingxi Zhang
- Xiao Liu
- Xianbao Zhan
- Zhaoshen Li
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Affiliations: Department of Gastroenterology, Changhai Hospital, Second Military Medical University, Shanghai 200433, P.R. China
Published online on: October 15, 2014 https://doi.org/10.3892/mmr.2014.2658
Pages: 3303-3309

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Abstract

The underlying malignancy of mucinous cystic neoplasms (MCNs) of the pancreas most commonly results in patients undergoing surgery. The tumorigenesis of MCNs remains elusive and few studies have investigated the role of specific micro (mi)RNAs in MCNs. The present study focused on the expression of miRNA‑224 and its putative target gene Jagged1 (Jag1) to examine its role in tumorigenesis and its suitability as a biomarker for MCNs. Paired tissue samples confirmed by surgical pathology were used to screen the miRNAs involved in MCNs with miRNA microarrays (n=3), and to verify the differentially expressed miRNAs (n=3) and mRNAs of candidate target genes of miRNAs by quantitative polymerase chain reaction (qPCR; n=8). Immunohistochemistry was conducted to confirm the expression and location of Jag1 in the neoplastic epithelial cells. Luciferase assays were performed to confirm the direct target gene of miRNA‑224. miRNA microarray analysis revealed that two differentially expressed miRNAs were closely associated with tumorigenesis and pancreatic diseases. The qPCR results revealed that miRNA‑224 was more significantly aberrantly expressed and the mRNA expression levels of its putative target gene, Jag1, were upregulated. Strong, diffuse cytoplasmic immunohistochemical labeling of Jag1 with occasional nuclear labeling was detected in the mucinous epithelium. Luciferase reporter activity was significantly reduced by co‑transfected miRNA‑224 mimics and pMIR‑Jag1‑wild-type, which suggested that miRNA‑224 bound to recognition sites in the 3' untranslated region of its target mRNA, Jag1. In conclusion, miRNA‑224 was downregulated in MCNs and may regulate tumorigenesis by targeting Jag1. Further studies investigating the role of miRNAs and functional analysis of epigenetic alterations are required to examine the diagnostic and therapeutic potential of miRNAs in MCNs.

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1	Crippa S, Fernández-Del CC, Salvia R, et al: Mucin-producing neoplasms of the pancreas: an analysis of distinguishing clinical and epidemiologic characteristics. Clin Gastroenterol Hepatol. 8:213–219. 2010. View Article : Google Scholar : PubMed/NCBI
2	Testini M, Gurrado A, Lissidini G, Venezia P, Greco L and Piccinni G: Management of mucinous cystic neoplasms of the pancreas. World J Gastroenterol. 16:5682–5692. 2010. View Article : Google Scholar : PubMed/NCBI
3	Grogan JR, Saeian K, Taylor AJ, Quiroz F, Demeure MJ and Komorowski RA: Making sense of mucin-producing pancreatic tumors. AJR Am J Roentgenol. 176:921–929. 2001. View Article : Google Scholar : PubMed/NCBI
4	Crippa S, Salvia R, Warshaw AL, et al: Mucinous cystic neoplasm of the pancreas is not an aggressive entity: lessons from 163 resected patients. Ann Surg. 247:571–579. 2008. View Article : Google Scholar : PubMed/NCBI
5	Matthaei H, Schulick RD, Hruban RH and Maitra A: Cystic precursors to invasive pancreatic cancer. Nat Rev Gastroenterol Hepatol. 8:141–50. 2011. View Article : Google Scholar
6	Goh BK, Tan YM, Chung YF, et al: A review of mucinous cystic neoplasms of the pancreas defined by ovarian-type stroma: clinicopathological features of 344 patients. World J Surg. 30:2236–2245. 2006. View Article : Google Scholar : PubMed/NCBI
7	Sawhney MS, Devarajan S, O’Farrel P, et al: Comparison of carcinoembryonic antigen and molecular analysis in pancreatic cyst fluid. Gastrointest Endosc. 69:1106–1110. 2009. View Article : Google Scholar : PubMed/NCBI
8	Bloomston M, Frankel WL, Petrocca F, et al: MicroRNA expression patterns to differentiate pancreatic adenocarcinoma from normal pancreas and chronic pancreatitis. JAMA. 297:1901–1908. 2007. View Article : Google Scholar : PubMed/NCBI
9	Szafranska AE, Doleshal M, Edmunds HS, et al: Analysis of microRNAs in pancreatic fine-needle aspirates can classify benign and malignant tissues. Clin Chem. 54:1716–1724. 2008. View Article : Google Scholar : PubMed/NCBI
10	Yu J, Li A, Hong SM, Hruban RH and Goggins M: MicroRNA alterations of pancreatic intraepithelial neoplasias. Clin Cancer Res. 18:981–992. 2012. View Article : Google Scholar : PubMed/NCBI
11	Remmers N, Bailey JM, Mohr AM and Hollingsworth MA: Molecular pathology of early pancreatic cancer. Cancer Biomark. 9:421–440. 2011.
12	du Rieu MC, Torrisani J, Selves J, et al: MicroRNA-21 is induced early in pancreatic ductal adenocarcinoma precursor lesions. Clin Chem. 56:603–612. 2010.
13	Jamieson NB, Morran DC, Morton JP, et al: MicroRNA molecular profiles associated with diagnosis, clinicopathologic criteria, and overall survival in patients with resectable pancreatic ductal adenocarcinoma. Clin Cancer Res. 18:534–545. 2012. View Article : Google Scholar
14	Habbe N, Koorstra JB, Mendell JT, et al: MicroRNA miR-155 is a biomarker of early pancreatic neoplasia. Cancer Biol Ther. 8:340–346. 2009. View Article : Google Scholar : PubMed/NCBI
15	Fukushima N, Sato N, Prasad N, Leach SD, Hruban RH and Goggins M: Characterization of gene expression in mucinous cystic neoplasms of the pancreas using oligonucleotide microarrays. Oncogene. 23:9042–9051. 2004. View Article : Google Scholar
16	Yao G, Yin M, Lian J, et al: MicroRNA-224 is involved in transforming growth factor-beta-mediated mouse granulosa cell proliferation and granulosa cell function by targeting Smad4. Mol Endocrinol. 24:540–551. 2010. View Article : Google Scholar : PubMed/NCBI
17	Izeradjene K, Combs C, Best M, et al: Kras(G12D) and Smad4/Dpc4 haploinsufficiency cooperate to induce mucinous cystic neoplasms and invasive adenocarcinoma of the pancreas. Cancer Cell. 11:229–243. 2007. View Article : Google Scholar : PubMed/NCBI
18	Mees ST, Mardin WA, Sielker S, et al: Involvement of CD40 targeting miR-224 and miR-486 on the progression of pancreatic ductal adenocarcinomas. Ann Surg Oncol. 16:2339–2250. 2009. View Article : Google Scholar : PubMed/NCBI
19	Rodilla V, Villanueva A, Obrador-Hevia A, et al: Jag1 is the pathological link between Wnt and Notch pathways in colorectal cancer. Proc Natl Acad Sci USA. 106:6315–6320. 2009. View Article : Google Scholar
20	Kiefel H, Bondong S, Erbe-Hoffmann N, et al: L1CAM-integrin interaction induces constitutive NF-kappaB activation in pancreatic adenocarcinoma cells by enhancing IL-1beta expression. Oncogene. 29:4766–4778. 2010. View Article : Google Scholar
21	Levy L and Hill CS: Smad4 dependency defines two classes of transforming growth factor {beta} (TGF-{beta}) target genes and distinguishes TGF-{beta}-induced epithelial-mesenchymal transition from its antiproliferative and migratory responses. Mol Cell Biol. 25:8108–8125. 2005. View Article : Google Scholar
22	Maniati E, Bossard M, Cook N, et al: Crosstalk between the canonical NF-κB and Notch signaling pathways inhibits Pparγ expression and promotes pancreatic cancer progression in mice. J Clin Invest. 121:4685–4699. 2011.
23	Wang Z, Banerjee S, Ahmad A, et al: Activated K-ras and INK4a/Arf deficiency cooperate during the development of pancreatic cancer by activation of Notch and NF-κB signaling pathways. PLoS One. 6:e205372011.PubMed/NCBI
24	Bao B, Wang Z, Ali S, et al: Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett. 307:26–36. 2011. View Article : Google Scholar : PubMed/NCBI
25	Vaccaro V, Melisi D, Bria E, et al: Emerging pathways and future targets for the molecular therapy of pancreatic cancer. Expert Opin Ther Targets. 15:1183–1196. 2011. View Article : Google Scholar : PubMed/NCBI
26	Prueitt RL, Yi M, Hudson RS, et al: Expression of microRNAs and protein-coding genes associated with perineural invasion in prostate cancer. Prostate. 68:1152–1164. 2008. View Article : Google Scholar : PubMed/NCBI
27	Davalos V, Moutinho C, Villanueva A, et al: Dynamic epigenetic regulation of the microRNA-200 family mediates epithelial and mesenchymal transitions in human tumorigenesis. Oncogene. 31:2062–2074. 2012. View Article : Google Scholar : PubMed/NCBI
28	Sorio C, Capelli P, Lissandrini D, et al: Mucinous cystic carcinoma of the pancreas: a unique cell line and xenograft model of a preinvasive lesion. Virchows Arch. 446:239–245. 2005. View Article : Google Scholar : PubMed/NCBI
29	Katoh M and Katoh M: Notch ligand, JAG1, is evolutionarily conserved target of canonical WNT signaling pathway in progenitor cells. Int J Mol Med. 17:681–685. 2006.PubMed/NCBI
30	Selcuklu SD, Donoghue MT, Kerin MJ and Spillane C: Regulatory interplay between miR-21, JAG1 and 17beta-estradiol (E2) in breast cancer cells. Biochem Biophys Res Commun. 423:234–239. 2012. View Article : Google Scholar : PubMed/NCBI
31	Pang RT, Leung CO, Ye TM, et al: MicroRNA-34a suppresses invasion through downregulation of Notch1 and Jag1 in cervical carcinoma and choriocarcinoma cells. Carcinogenesis. 31:1037–1044. 2010. View Article : Google Scholar : PubMed/NCBI
32	Steg AD, Katre AA, Goodman B, et al: Targeting the notch ligand JAGGED1 in both tumor cells and stroma in ovarian cancer. Clin Cancer Res. 17:5674–5685. 2011. View Article : Google Scholar : PubMed/NCBI
33	Brabletz S, Bajdak K, Meidhof S, et al: The ZEB1/miR-200 feedback loop controls Notch signalling in cancer cells. EMBO J. 30:770–782. 2011. View Article : Google Scholar : PubMed/NCBI
34	Ban J, Bennani-Baiti IM, Kauer M, et al: EWS-FLI1 suppresses NOTCH-activated p53 in Ewing’s sarcoma. Cancer Res. 68:7100–7109. 2008.PubMed/NCBI
35	Sasaki Y, Ishida S, Morimoto I, et al: The p53 family member genes are involved in the Notch signal pathway. J Biol Chem. 277:719–724. 2002. View Article : Google Scholar : PubMed/NCBI
36	Dickson BC, Mulligan AM, Zhang H, et al: High-level JAG1 mRNA and protein predict poor outcome in breast cancer. Mod Pathol. 20:685–693. 2007. View Article : Google Scholar : PubMed/NCBI
37	Golson ML, Le Lay J, Gao N, et al: Jag1 is a competitive inhibitor of Notch signaling in the embryonic pancreas. Mech Dev. 126:687–699. 2009. View Article : Google Scholar : PubMed/NCBI
38	Bao B, Wang Z, Ali S, et al: Notch-1 induces epithelial-mesenchymal transition consistent with cancer stem cell phenotype in pancreatic cancer cells. Cancer Lett. 307:26–36. 2011. View Article : Google Scholar : PubMed/NCBI
39	Golson ML, Loomes KM, Oakey R and Kaestner KH: Ductal malformation and pancreatitis in mice caused by conditional Jag1 deletion. Gastroenterology. 136:1761–1771. 2009. View Article : Google Scholar : PubMed/NCBI
40	Fukushima N, Sato N, Prasad N, Leach SD, Hruban RH and Goggins M: Characterization of gene expression in mucinous cystic neoplasms of the pancreas using oligonucleotide microarrays. Oncogene. 23:9042–9051. 2004. View Article : Google Scholar : PubMed/NCBI
41	Tan MH, Nowak NJ, Loor R, et al: Characterization of a new primary human pancreatic tumor line. Cancer Invest. 4:15–23. 1986. View Article : Google Scholar : PubMed/NCBI
42	Volinia S, Calin GA, Liu CG, et al: A microRNA expression signature of human solid tumors defines cancer gene targets. Proc Natl Acad sci USA. 103:2257–2261. 2006. View Article : Google Scholar : PubMed/NCBI
43	Zhang S, Hao J, Xie F, Hu X, et al: Downregulation of miR-132 by promoter methylation contributes to pancreatic cancer development. Carcinogenesis. 32:1183–1189. 2011. View Article : Google Scholar : PubMed/NCBI